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The expression of viral genes during the productive life cycle of human papillomaviruses (HPV) is tightly coupled to the differentiation program of epithelial cells. We have examined transcription of HPV as a function of differentiation in an in vitro organotypic raft culture system which allows for epithelial stratification at the air-liquid interface. When CIN612 cells, which contain episomal copies of HPV type 31b (HPV31b), were allowed to stratify in raft cultures, they differentiated in a manner which was histologically similar to that seen in a cervical intraepithelial neoplasia I biopsy lesion. In monolayer cultures of CIN612 cells, two major polycistronic HPV31b transcripts of 1.7 kb which encode (i) E6, E7, E1-E4, and E5 and (ii) E6*, E7, E1-E4, and E5 were identified. These RNAs initiated at a promoter, P97, in the upstream regulatory region of the virus. Following differentiation in raft cultures, the relative abundance of RNAs initiated at P97 was unchanged. In contrast, the expression of a 1.3-kb RNA encoding an E1-E4 fusion protein and E5 was found to increase substantially following differentiation. This transcript was initiated at a novel promoter within the E7 gene (P742). These studies have therefore identified a constitutive viral promoter which is active throughout stratified epithelium as well as a novel promoter which is induced upon epithelial cell differentiation.

Differentiation-induced and constitutive transcription of human papillomavirus type 31b in cell lines containing viral episomes